LOCUS       sequence-242487-        6273 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6273)
  AUTHORS   Yang J, Sanderson NS, Wawrowsky K, Puntel M, Castro MG, Lowenstein 
            PR
  TITLE     Kupfer-type immunological synapse characteristics do not predict 
            anti-brain tumor cytolytic T-cell function in vivo.
  JOURNAL   Proc Natl Acad Sci U S A. 2010 Mar 9. 107(10):4716-21.
  PUBMED    20133734
REFERENCE   2  (bases 1 to 6273)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 2010 Mar 9. 107(10):4716-21."
COMMENT     Sequence Label: pCI-neo-cOVA
FEATURES             Location/Qualifiers
     source          1..6273
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        79..458
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        459..670
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     621..641
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     667..691
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          831..963
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     946..965
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1008..1026
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1442..1465
                     /codon_start=1
                     /product="antigenic CD8+ T cell epitope from chicken 
                     ovalbumin"
                     /label=OVA peptide
                     /translation="SIINFEKL"
     polyA_signal    1918..2039
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1929..1948)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1983..2002
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2225..2680
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2312..2331)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2522..2543
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2797..3154
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2797..2988
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2820..2840)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3005..3140
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3205..3999
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3259..3278)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3869..3888
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4063..4111
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4152..4171)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4271..4293
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4331..4349)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4417..4521
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4522..4590,4591..5382)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4740..4759)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5553..6141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6042..6061
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
       61 caatatgacc gccatgttgg cattgattat tgactagtta ttaatagtaa tcaattacgg
      121 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      181 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      241 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      301 cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg
      361 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt
      421 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      481 ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      541 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      601 gcgatcgccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      661 aagcagagct cgtttagtga accgtcagat cactagaagc tttattgcgg tagtttatca
      721 cagttaaatt gctaacgcag tcagtgcttc tgacacaaca gtctcgaact taagctgcag
      781 tgactctctt aaggtagcct tgcagaagtt ggtcgtgagg cactgggcag gtaagtatca
      841 aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga cagagaagac
      901 tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc tttctctcca
      961 caggtgtcca ctcccagttc aattacagct cttaaggcta gagtacttaa tacgactcac
     1021 tataggctag cctcgagaat tcgtcgacat gggctccatc ggcgcagcaa gcatggaatt
     1081 ttgttttgat gtattcaagg agctcatcaa ttcctgggta gaaagtcaga caaatggaat
     1141 tatcagaaat gtccttcagc caagctccgt ggattctcaa actgcaatgg ttctggttaa
     1201 tgccattgtc ttcaaaggac tgtgggagaa aacatttaag gatgaagaca cacaagcaat
     1261 gcctttcaga gtgactgagc aagaaagcaa acctgtgcag atgatgtacc agattggttt
     1321 atttagagtg gcatcaatgg cttctgagaa aatgaagatc ctggagcttc catttgccag
     1381 tgggacaatg agcatgttgg tgctgttgcc tgatgaagtc tcaggccttg agcagcttga
     1441 gagtataatc aactttgaaa aactgactga atggaccagt tctaatgtta tggaagagag
     1501 gaagatcaaa gtgtacttac ctcgcatgaa gatggaggaa aaatacaacc tcacatctgt
     1561 cttaatggct atgggcatta ctgacgtgtt tagctcttca gccaatctgt ctggcatctc
     1621 ctcagcagag agcctgaaga tatctcaagc tgtccatgca gcacatgcag aaatcaatga
     1681 agcaggcaga gaggtggtag ggtcagcaga ggctggagtg gatgctgcaa gcgtctctga
     1741 agaatttagg gctgaccatc cattcctctt ctgtatcaag cacatcgcaa ccaacgccgt
     1801 tctcttcttt ggcagatgtg tttcccctta agtcgacgaa ttcacgcgtg gtacctctag
     1861 agtcgacccg ggcggccgct tccctttagt gagggttaat gcttcgagca gacatgataa
     1921 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     1981 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     2041 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt
     2101 aaagcaagta aaacctctac aaatgtggta aaatccgata aggatcgatc cgggctggcg
     2161 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     2221 atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2281 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2341 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2401 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2461 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2521 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2581 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2641 tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     2701 ttacgcatct gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac catggcctga
     2761 aataacctct gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg
     2821 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2881 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2941 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3001 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3061 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     3121 aggaggcttt tttggaggcc taggcttttg caaaaagctt gattcttctg acacaacagt
     3181 ctcgaactta aggctagagc caccatgatt gaacaagatg gattgcacgc aggttctccg
     3241 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3301 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3361 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3421 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3481 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3541 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3601 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3661 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3721 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3781 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3841 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3901 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3961 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4021 tgaccgacca agcgacgccc aacctgccat cacgatggcc gcaataaaat atctttattt
     4081 tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat ccgcgtatgg
     4141 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4201 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4261 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4321 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4381 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4441 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4501 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4561 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4621 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4681 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4741 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4801 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4861 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4921 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4981 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5041 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5101 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5161 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5221 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5281 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5341 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5401 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5461 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5521 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5581 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5641 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5701 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5761 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5821 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5881 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5941 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6001 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6061 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6121 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6181 tgctggcctt ttgctcacat ggctcgacag atcttcaata ttggccatta gccatattat
     6241 tcattggtta tatagcataa atcaatattg gct
//
